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Objective:To compare ocular surface dry eye-related indexes and tear cytokine level changes in chronic ocular graft-versus-host disease (oGVHD) patients after receiving topical treatment of 0.05% cyclosporine or 0.1% tacrolimus eye drops.Methods:A randomized controlled study was conducted.A total of 60 chronic oGVHD patients (60 eyes) were recruited at Beijing University Third Hospital from April 2020 to April 2021.The patients were divided into tacrolimus group and cyclosporine group by a random number table, with 30 patients (30 eyes) in each group.Patients in tacrolimus group used 0.1% tacrolimus eye drops (twice a day) and patients in cyclosporine group used 0.05% cyclosporine eye drops (4 times a day).Additionally, 0.1% flumetholon (twice a day), deproteinized calf blood extract (3 times a day), and 0.1% sodium hyaluronate eye drops (8 times a day) were applied for anti-inflammation and lubrication in both groups.Patients were screened according to exclusion criteria after 1-month treatment.Eventually, 21 patients (21 eyes) in tacrolimus group and 12 patients (12 eyes) in cyclosporine group were included for further study.Patients were examined before and 1 month after treatment.The primary evaluation indexes included Ocular Surface Disease Index (OSDI), corneal fluorescein staining scores and tear film break-up time (BUT).Expressions of interleukin (IL)-6, IL-8, IL-17, epidermal growth factor (EGF), and tumor necrosis factor-α (TNF-α) in tears were detected before and after treatment using Luminex chip.This study adhered to the Declaration of Helsinki.The study protocol was approved by the Ethics Committee of Peking University Third Hospital (No.M2020489).Written informed consent was obtained from each subject before any medical examination.Results:The OSDI differences between before and after treatment were 0.4(-5.6, 2.5) in tacrolimus group and 27.2(4.6, 45.0) in cyclosporine group, and the OSDI improvement was significantly greater in cyclosporine group than in tacrolimus group ( Z=-2.547, P=0.009).The differences of corneal fluorescein staining scores and BUT between before and after treatment were 5.0(2.5, 10.0) scores and 3.5(-0.5, 13.8) seconds in tacrolimus group, 0.0(-3.0, 0.0) scores and -1.5(-3.0, 0.0) seconds in cyclosporine group, respectively, with no significantly difference between both groups ( Z=-0.526, -0.804; both at P>0.05).The differences of IL-6, IL-8, IL-17, EGF and TNF-α expressions between before and after treatment in tacrolimus group and cyclosporine group were not significantly different ( Z=-0.487, -0.112, -0.412, -1.085, -1.198; all at P>0.05). Conclusions:Altered levels of all tested cytokines in oGVHD tears are of no significant differences between tacrolimus and cyclos porine treatment.In addition, 0.05% cyclosporine eye drops may be more comfortable than 1% tacrolimus for chronic oGVHD patients.
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Objective:To investigate the virological testing results of patients with corneal graft at the time of repeat keratoplasty and the diagnostic efficacy of multiple viral examinations.Methods:A case-control study was conducted.A total of 14 consecutive patients diagnosed with corneal graft failure were enrolled as graft failure group from March 2018 to December 2018 in Peking University Third Hospital, and 15 consecutive patients diagnosed with bullous keratopathy (BK) were enrolled as BK group in the meantime.All patients had unilateral involvement and indications for keratoplasty.Serum samples were collected from venous blood on the day of surgery, and specimens of aqueous humor and corneal tissue were obtained during corneal transplantation.Viral DNA in aqueous humor and corneal specimens was detected by real-time polymerase chain reaction (PCR).The level of viral antibodies in serum and aqueous humor was determined by enzyme-linked immunosorbent assay and the Goldmann-Witmer coefficient (GWC) was calculated.The tested viral species included herpes simplex virus (HSV), herpes zoster virus (VZV), and cytomegalovirus (CMV).For graft failure group, the relevance between elevated intraocular pressure, multiple previous keratoplasty surgeries, histories of viral keratitis and any positive result of viral analyses in this study were measured by the kappa consistency test.This study adhered to the Declaration of Helsinki.The study protocol was approved by the Ethics Committee of Peking University Third Hospital (No.2017299-02).Written informed consent was obtained from each subject.Results:At the time of keratoplasty, 9 out of 14 eyes in the graft failure group tested positive for at least one type of virus, with 6 eyes positive for CMV and 3 eyes positive for VZV.Positive aqueous humor PCR analysis detected VZV in 5 out of 14 eyes.Corneal tissue PCR analysis detected CMV in 5 out of 14 eyes.Positive GWC calculations detected CMV in 3 out of 10 eyes.The concordance between viral DNA and antibody detection was poor.All eyes in BK group were negative for the virological test, except for 2 eyes (2/15) with elevated aqueous humor GWC for CMV.The prevalence of viral infection was 64.2%(9/14) in graft failure group, which was significantly higher than 13.3%(2/15) in BK group ( P=0.014).In graft failure group, 7 eyes (7/14) had elevated intraocular pressure, 3 eyes (3/14) had multiple keratoplasty surgeries, and 6 eyes (6/14) had viral keratitis before this keratoplasty.However, none of these factors showed significant relevance with positive virological results (kappa=0.143, -0.155, -0.286). Conclusions:Viral infection has become a major cause of corneal graft failure.A combination of various virological analyses during keratoplasty can help to clarify the etiology and the viral infection status, and ultimately guide subsequent treatment.
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Objective:To investigate the distribution of herpesvirus DNA in the corneal layers of herpesvirus-positive corneal transplantation patients and to compare the efficiency of viral DNA detection in corneal and aqueous humor samples in these patients.Methods:A diagnostic test study was conducted.Data from patients, who underwent keratoplasty in Peking University Third Hospital from May 2015 to August 2021 and tested positive for herpesvirus in corneal tissue and/or aqueous humor samples obtained during surgery, were collected through the clinical medical record system.The demographic data and virus distribution layers of these patients were analyzed.The rate of herpesvirus detection in different samples was analyzed.The sensitivity of different samples for viral DNA detection was analyzed by receiver operating characteristic curves, and area under the curve (AUC) and 95% confidence interval ( CI) were recorded.This study adhered to the Declaration of Helsinki and the study protocol was approved by the Ethics Committee of Peking University Third Hospital (No.M2021283).Written informed consent was obtained from each patient before entering the cohort. Results:A total of 166 herpesvirus-positive patients (166 eyes) were collected.Of the 166 eyes, 75 eyes (45.2%) were positive for cytomegalovirus (CMV), 34 eyes (20.5%) for herpes simplex virus 1 (HSV-1), 30 eyes (18.1%) for varicella zoster virus (VZV), and 27 eyes (16.3%) for Epstein-Barr virus (EBV).CMV DNA and VZV DNA were detected in the endothelial layers of 47 eyes (62.7%) and 26 eyes (86.7%), respectively, which were significantly higher than the 28 eyes (37.3%) and 4 eyes (13.3%) with virus located in stromal layers ( χ2=4.813, 16.133; both at P<0.05).HSV-1 DNA and EBV DNA were detected in the endothelial layer of 8 eyes (23.5%) and 5 eyes (18.5%), respectively, which were less than the 26 eyes (76.5%) and 22 eyes (81.5%) with virus located in stromal layers, and the differences were statistically significant ( χ2=9.529, 10.704; both at P<0.001).The sensitivity of corneal samples for herpesvirus DNA positivity was 71.6%, which was higher than 54.1% of aqueous humor.The detection sensitivities of corneal samples for CMV DNA and VZV DNA positivity were 64.3%(AUC=0.821, 95% CI: 0.705-0.938) and 35.7%(AUC=0.679, 95% CI: 0.475-0.882), respectively, which were lower than 71.4%(AUC=0.875, 95% CI: 0.750-0.964) and 85.7%(AUC=0.929, 95% CI: 0.816-1.000) of aqueous humor samples.The detection sensitivities of corneal samples for HSV-1 DNA and EBV DNA were 100%(AUC=1.000, 95% CI: 1.000-1.000) and 92.3%(AUC=0.962, 95% CI: 0.875-1.000), respectively, which were higher than 27.8%(AUC=0.639, 95% CI: 0.455-0.823) and 23.1%(AUC=0.615, 95% CI: 0.395-0.835) of aqueous humor samples. Conclusions:The detection rate of CMV DNA is highest among herpesvirus-positive keratoplasty patients.CMV DNA and VZV DNA are primarily located in the corneal endothelial layers, while HSV-1 DNA and EBV DNA are more predominant in the corneal stromal layer.The sensitivity of virus DNA detection is higher in the cornea than in aqueous humor.
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Objective:To explore the long-term influence of donor central graft thickness (CGT) and donor graft size on corneal endothelial cell density (ECD) after Descemet stripping automated endothelial keratoplasty (DSAEK).Methods:An observational case series study was conducted.One hundred and forty-four eyes of 134 patients who underwent DSAEK in Peking University Third Hospital from January 2013 to December 2017 with at least 24-month follow-up were enrolled.Preoperative donor ECD was evaluated by specular microscopy, and ECD was determined by in vivo confocal microscopy at 1, 3, 6, 12, and 24 months postoperatively.Donor CGT was measured by anterior segment optical coherence tomography.According to the 3-month postoperative donor CGT, the subjects were divided into thinner graft group (45 eyes with CGT<100 μm), medium-thick graft group (66 eyes with CGT≥100-<150 μm) and thicker graft group (33 eyes with CGT≥150 μm). According to the donor trephination size, the subjects were divided into smaller graft group (31 eyes with trephination size≥7-<8 mm) and larger graft group (113 eyes with trephination size≥8-<9 mm). The changes of the donor CGT and corneal endothelial cell loss rate were compared at different time points after surgery.The relationships between 24-month postoperative ECD and donor ECD, donor graft size and donor CGT were analyzed.This study adhered to the Declaration of Helsinki.The study protocol was approved by the Ethics Committee of Peking University Third Hospital (No.IRB00006761-2008025). Written informed consent was obtained from each subject prior to any medical examination. Results:The donor CGT was 129.0 (90.8, 160.8), 115.5 (93.0, 146.0), 115.5 (89.0, 151.0), 112.5 (94.3, 146.8) and 114.0 (89.0, 144.5) μm at 1, 3, 6, 12 and 24 months after surgery, showing a statistically significant difference ( H=37.369, P<0.001). There was a statistically significant difference between 1-month and 3-month postoperative CGT ( P<0.001). There was no statistically significant difference in the endothelial cell loss rate among the three different donor CGT groups and between the two different donor graft size groups at any postoperative time points (all at P>0.05). Spearman correlation analysis showed that the 24-month postoperative ECD was strongly positively correlated with the preoperative donor ECD( rs=0.783, P<0.001), which was not associated with donor graft size and donor CGT ( rs=0.141, P=0.093; rs=-0.044, P=0.600). Conclusions:Larger postoperative ECD is correlated with larger preoperative ECD of donor graft.Lower long-term corneal endothelial cell loss rate after DSAEK is associated with thinner and larger diameter of donor graft.
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<p><b>BACKGROUND</b>Though there have been various methods for harvesting and preserving descemet membrane (DM) and intact endothelium, there is no literature about the morphological evaluation of endothelium after graft preparation for descemet membrane endothelial keratoplasty (DMEK). The aim of this study was to establish and improve a simple method for preparing, preserving, and morphologically evaluating the donor graft for DMEK.</p><p><b>METHODS</b>To obtain a donor graft, an air bubble was formed by injecting a 29 G needle with 1 ml sterile air into a small edge created outside the Schwalbe line. Another needle was inserted into the bubble through the stroma to aspirate the air or replace half the air with organ culture medium. Trypan blue was used to mark the location for small incision to improve the success rate. Frozen sections were stained with hematoxylin and eosin (HE). Based on the air bubble, DM grafts were divided into four groups: group A (normal control), graft without any operative technique; group B, graft with zero-pressure air bubble; group C, graft with full-pressure air bubble; group D, graft with half-pressure air bubble. The four groups of grafts were preserved for 24 hours to observe the effect of bubbles on cells. The gross and ultrastructure morphologies were evaluated using alizarin red and scanning electron microscopy (SEM), respectively.</p><p><b>RESULTS</b>Donor grafts were harvested via the air bubble technique, facilitated by prior trypan blue staining. HE-stained sections revealed a pure graft without stroma. There were no significant changes under light microscope. In group A, SEM revealed a confluent layer of polygonal endothelium with distributed microvilli exhibiting characteristics of interdigitating junctions. In group B, intercellular borders became thinner. In group C, interdigitations were almost flat and microvilli were observed less frequently. In group D, other than less microvilli, there were minimal changes.</p><p><b>CONCLUSIONS</b>The donor graft preparation method appears to be effective and convenient. Properly decreasing the air pressure could protect and preserve the endothelium.</p>
Subject(s)
Animals , Rabbits , Descemet Membrane , Cell Biology , Descemet Stripping Endothelial Keratoplasty , Methods , Endothelium, Corneal , Cell Biology , Tissue DonorsABSTRACT
Background Descemet stripping automated endothelial keratoplasty (DSAEK) is the main treatment for corneal endothelial dysfunction.But the visual outcome after operation varies depending on the difference of corneal diseases.Objective This study was to evaluate and compare the visual outcomes following DSAEK in different keratopathy.Methods The clinical data of 72 eyes of 67 patients with endothelial dysfunction underwent DSAEK from December 2007 to December 2009 at Peking University Third Hospital were retrospectively analyzed.The patients were divided into Fuchs endothelial dystrophy (FED) group (22 eyes of 19 cases),cataract surgeryinduced bullous keratopathy group (37 eyes of 36 eases) and other endothelial keratopathy group (12 eyes of 12 cases).The distribution of visual acuity and LogMAR acuities were compared and evaluation among the 3 groups before operation and 1 day,3,7,30,90 and 180 days after operation.Results Preoperatively,the vision was lower than 0.1 in 71.83％ patients,and all the patients had the visual acuity less than 0.3.No significant difference was found in the preoperative acuity among the FED group,cataract surgery-induced bullous keratopathy group and other endothelial keratopathy group(x2 =3.427,P＞0.05).The visual acuity was ≥ 0.4 in 40 eyes (56.34％) and ≥ 0.1 in 65 eyes (91.55％),and the percentage of vision ≥0.8 was 22.73％ in the FED group,10.81％ in the cataract surgery-induced bullous keratopathy group and 8.33％ in the other endothelial keratopathy group,and no significant difference was found in the percentage of eyes in different visual acuities 180 days after operation (x2 =0.330,P＞ 0.05).In the 3 groups,LogMAR values were gradually decreased with the lapse of the time,showing a significant difference (Ftime =88.000,P ＜ 0.01).In the seventh day after operation,LogMAR value was 1.29 ± 0.57 in the cataract surgery-induced bullous keratopathy group,which was significantly higher than that of the FED group (0.82± 0.43) or other endothelial keratopathy group (0.91 ±0.39) (both at P＜0.05).Ninety days after operation,LogMAR value was 0.40 ±0.28 in the FED group and was significantly declined in comparison with the cataract surgeryinduced bullous keratopathy group (0.64±0.44) and other endothelial keratopathy group (0.73±0.54) (both at P＜0.05).However,no significant differences was seen in the LogMAR values among the three groups 180 days after operation (all at P＞0.05).The vision was stable 3 months after operation in the FED group,however,the vision was still changed over 3 months in the cataract surgery-induced bullous keratopathy group and other endothelial keratopathy group.Conclusions DSAEK is available for corneal endothelial diseases.Visual acuity improves more rapid in FED patients than other keratopathies.